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原阿片碱对人肝星状细胞HSC-LX2增殖的抑制作用及机制研究
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| 篇名： | 原阿片碱对人肝星状细胞HSC-LX2增殖的抑制作用及机制研究 |
| TITLE： | |
| 摘要： | 目的：考察原阿片碱对人肝星状细胞HSC-LX2增殖的抑制作用，并对其机制进行初步探讨。方法：采用MTT法测定25、50、100、200、400、500 μmol/L原阿片碱作用24 h对HSC-LX2细胞增殖的影响，计算细胞增殖抑制率。另取HSC-LX2细胞分为对照组（含5％胎牛血清的1640培养基）和原阿片碱低、中、高浓度组（100、200、400 μmol/L），作用24 h后，流式细胞术测定细胞凋亡率；荧光定量逆转录-聚合酶链式反应（RT-PCR）法测定细胞中α-平滑肌肌动蛋白（α-SMA）、Ⅰ型胶原蛋白（Collagen Ⅰ）、Ⅲ型胶原蛋白（Collagen Ⅲ）、基质金属蛋白酶2（MMP-2）、组织金属蛋白酶抑制剂1（TIMP-1） mRNA的相对表达量，Western blot法测定细胞中α-SMA、Collagen Ⅰ、Collagen Ⅲ、MMP-2蛋白的相对表达量。结果：25、50、100、200、400、500 μmol/L原阿片碱对HSC- LX2细胞的增殖抑制率分别为0、6.9％、18.7％、34.2％、48.9％、53.9％。与对照组比较，原阿片碱低、中、高浓度组细胞中Collagen Ⅰ、TIMP-1 mRNA相对表达量和α-SMA蛋白相对表达量均显著降低，MMP-2蛋白相对表达量显著升高，差异均有统计学意义（P＜0.05或P＜0.01）；原阿片碱中、高浓度组细胞的凋亡率和MMP-2 mRNA相对表达量显著升高，α-SMA、Collagen Ⅲ mRNA相对表达量和Collagen Ⅰ、Collagen Ⅲ 蛋白相对表达量均显著降低，差异均有统计学意义（P＜0.05或P＜0.01）。 结论：原阿片碱可抑制HSC-LX2细胞增殖，诱导其凋亡，可降低α-SMA、Collagen Ⅰ、Collagen Ⅲ、TIMP-1表达，升高MMP-2表达有关。 |
| ABSTRACT： | OBJECTIVE： To investigate inhibitory effects of protopine on the proliferation of human hepatic stellate cells HSC-LX2 and to explore its mechanism preliminarily. METHODS： MTT assay was used to detect the effects of 25， 50， 100， 200， 400 and 500 μmol/L protopine on the proliferation of HSC-LX2 cells. The inhibitory effect of cell proliferation was calculated. HSC-LX2 cells were divided into control group （1640 medium containing 5％ fetal bovine serum）， protopine low-concentration， medium-concentration and high-concentration groups （100， 200， 400 μmol/L）. After treated for 24 h. The apoptotic rate of the cells was detected by flow cytometry. RT-PCR was used to determine the mRNA expression of α-SMA， Collagen Ⅰ， Collagen Ⅲ， MMP-2 and TIMP-1 in cells. The protein expressions of α-SMA， Collagen Ⅰ， Collagen Ⅲ and MMP-2 were detected by Western blot. RESULTS： The inhibitory rates of 25， 50， 100， 200， 400 and 500 μmol/L protopine on proliferation HSC-LX2 cells were 0， 6.9％， 18.7％， 34.2％， 48.9％， 53.9％， respectively. Compared with control group， mRNA expression of Collagen Ⅰ， TIMP-1 and protein expression of α-SMA were decreased significantly in protopine low-concentration， medium-concentration and high-concentration groups， while protein expression of MMP- 2 was increased significantly， with statistical significance （P＜0.05 or P＜0.01）. Apoptotic rate of HSC-LX2 cells and mRNA expression of MMP-2 were increased significantly in protopine medium-concentration and high-concentration groups， mRNA expression of α-SMA and Collagen Ⅲ， protein expression of Collagen Ⅰ and Collagen Ⅲ were decreased significantly， with statistical significance （P＜0.05 or P＜0.01）. CONCLUSIONS： Protopine can induce the apoptosis of HSC-LX2 cells and inhibit their cell proliferation， and reduce the expression of a-SMA， Collagen Ⅰ， Collagen Ⅲ and TIMP-1， and increase the expression of MMP-2. |
| 期刊： | 2019年第30卷第23期 |
| 作者： | 吴新玉，李靖，张金娟，廖尚高，梅青，吴亚云，席晓岚 |
| AUTHORS： | WU Xinyu，LI Jing，ZHANG Jinjuan，LIAO Shanggao，MEI Qing，WU Yayun，XI Xiaolan |
| 关键字： | 原阿片碱；肝星状细胞HSC-LX2；增殖；凋亡；机制 |
| KEYWORDS： | Protopine； Hepatic stellate cells HSC-LX2； Proliferation； Apoptosis； Mechanism |
| 阅读数： | 718 次 |
| 本月下载数： | 6 次 |
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